Thrombelastography in 26 healthy horses with and without activation by recombinant human tissue factor

Objectives – To develop a standardized technique for thrombelastography (TEG) analysis in healthy adult horses, with and without the ex vivo addition of tissue factor (TF) as an activator. To determine reference intervals for TEG parameters in the horse, and to determine if traditional coagulation tests correlate with TEG. Design – Prospective, observational. Setting – Veterinary teaching hospital. Animals – Twenty‐six healthy adult horses. Interventions – None. Measurements and Main Results – Thrombelastography with (TF‐TEG) and without (TEG) the addition of TF performed by 4 operators. Coagulation profiles (prothrombin time, activated partial thromboplastin time, platelet count, fibrinogen, antithrombin, and fibrinogen degradation products) were assessed in a subset of horses. Mean values (SD) for TEG parameters in healthy horses were: reaction time (R)=17.0 minutes (3.0 min), K time (K)=5.8 minutes (2.3 min), clotting rate (Ang)=42° (14°), maximum clot strength (maximum amplitude [MA])=60.3 mm (5.7 mm), CL30=97.0% (2.0%), LY30=0.8% (0.6%), CL60=92% (5.9%), LY60=3.2% (2.5%). Mean values (SD) for TF‐TEG parameters were: R‐TF=6.6 minutes (1.4 min), K‐TF=3.1 minutes (1.0 min), Ang‐TF=50.9° (9°), MA‐TF=62.3 mm (5.1 mm), CL30‐TF=97.8% (1.6%), LY30‐TF=0.6% (0.5%), CL60‐TF=90.8% (4.2%), and LY60‐TF=3.6% (1.9%). The addition of TF decreased R and K and increased Ang. TF‐TEG had a narrower SD for R, K, Ang, CL60 and LY60 compared with TEG. Interoperator differences were reduced by the addition of TF. Regression analysis indicated a positive relationship between MA and fibrinogen concentrations (P=0.02) and R‐TF time and prothrombin time (P=0.03). Conclusion – TF‐TEG using the described protocol may minimize variability in data obtained across institutions or users. However, due to the variability associated with different operators, it is recommended that each laboratory set up individual reference intervals with the personnel who will perform the assay, and that the assay protocols and data obtained are compared on a regular basis.     

Association of admission plasma D-dimer concentration with diagnosis and outcome in horses with colic

Background: Coagulopathies detected in horses with gastrointestinal problems seem to be associated with poor outcome. Plasma D‐Dimer concentration is a sensitive test for assessing coagulopathies. Hypothesis: Plasma D‐Dimer concentration tested on admission is related to diagnosis and outcome in horses with colic. Animals: Four hundred and ninety three horses referred for evaluation of abdominal pain. Methods: Prospective observational clinical study. Horses were grouped according to diagnosis (medical and surgical intestinal obstructions, ischemic disorders with and without intestinal resection, enteritis, peritonitis), outcome (survivors, nonsurvivors), and number of coagulopathies (normal profile, 1 or 2 coagulopathies, subclinical disseminated intravascular coagulation [DIC]). Blood samples were collected on admission and plasma D‐Dimer concentration, clotting times (PT and aPTT), and antithrombin activity were determined. Positive likelihood ratios (LR+) were calculated for evaluation of D‐Dimer cut‐off values, which were later tested in a logistic regression model. Results: Horses with enteritis or peritonitis had significantly (P < .001) higher plasma D‐Dimer concentrations and more severe coagulopathies on admission than horses with other diagnoses. Nonsurvivors also had significantly (P < .001) higher plasma D‐Dimer concentrations at presentation than did survivors, and those horses with subclinical DIC on presentation had an odds ratio (OR) 8.6 (95% confidence interval [CI], 3.3–22.5, P < .001) for nonsurvival. Finally, D‐Dimer concentrations >4,000 ng/mL had a LR+ of 5.9 and an OR 8.8 (95% CI, 4.5–17.1, P < .001) for nonsurvival. Conclusion and Clinical Importance: Plasma D‐Dimer concentration measured on admission can be used to facilitate diagnosis and outcome prediction in horses with colic. A potential cut‐off value for nonsurvival was found at approximately 4,000 ng/mL.     

Thrombelastography in horses with acute gastrointestinal disease

Background: Coagulopathies in horses with gastrointestinal disease are frequently identified and associated with morbidity and fatality. Objective: Determine if thrombelastography (TEG) identifies abnormalities associated with lesion type, presence of systemic inflammatory response syndrome (SIRS), morbidity, and fatality more consistently than traditional coagulation testing. Animals: One‐hundred and one horses examined for gastrointestinal disease and 20 healthy horses. Methods: TEG, tissue factor (TF)‐TEG, and traditional coagulation panels parameters and percentages of horses with coagulopathies were compared for lesion type, presence of SIRS, complications, and survival. Results: Changes in individual parameters and increased incidence of coagulopathies were associated with fatality (R, P= .007; k‐value [K], P= .004; clot lysis [CL]30, P= .037; CL60, P= .050; angle [Ang], P= .0003; maximum amplitude [MA], P= .006; lysis [Ly]30, P= .042; Ly60, P= .027; CI, P= .0004; ≥ 2 TEG coagulopathies, P= .013; ≥ 3 TEG coagulopathies, P= .038; TF‐R, P= .037; TF‐K, P= .004; TF‐CL30, P < .0001; TF‐CL60, P < .0001; TF‐Ang, P= .005; TF‐Ly30, P= .0002; TF‐Ly60, P < .0001; TF‐CI, P= .043; ≥ 1 TF‐TEG coagulopathies, P= .003; ≥ 2 TF‐TEG coagulopathies, P= .0004; prothrombin tme [PT], P < .0001; activated partial throboplastin time [aPTT], P= .021), inflammatory lesions (MA, P= .013; TF‐CL30, P= .033; TF‐CL60, P= .010; TF‐Ly60, P= .011; ≥ 1 TF‐TEG coagulopathy, P= .036; ≥ 2 TF‐TEG coagulopathy, P= .0007; PT, P= .0005; fibrinogen, P= .019), SIRS (MA, P= .004; TF‐CL30, P= .019; TF‐CL60, P= .013; TF‐Ly30, P= .020; TF‐Ly60, P= .010; PT, P < .0001; aPTT, P= .032; disseminated intravascular coagulation, P= .005), and complications (ileus: aPTT, P= .020; diarrhea: TF‐CL30, P= .040; TF‐Ly30, P= .041; thrombophlebitis: ≥ 1 TF‐TEG coagulopathy, P= .018; laminitis: MA, P= .004; CL60, P= .045; CI, P= .036; TF‐MA, P= .019; TF‐TEG CI, P= .019). Abnormalities in TEG and TF‐TEG parameters were indicative of hypocoagulation and hypofibrinolysis. Conclusions and Clinical Importance: TEG identifies changes in coagulation and fibrinolysis associated with lesion type, SIRS, morbidity, and fatality in horses with gastrointestinal disease.     

Evaluation of a modified thrombelastography assay initiated with recombinant human tissue factor in clinically healthy horses

Background: Thrombelastography (TEG) is used to evaluate the viscoelastic properties of blood during clotting and provides a global assessment of hemostasis and clot lysis. TEG analysis initiated with recombinant human tissue factor (TF) has not been evaluated in clinically healthy horses. Objectives: The purpose of this study was to determine whether TEG results are affected by the time elapsed between sampling and analysis (storage time) of equine blood samples and to establish a preliminary equine reference interval for a modified TEG assay, using recombinant human TF to initiate coagulation. Methods: Citrated blood samples were obtained from 20 clinically healthy adult horses. Thirteen samples were stored for 30, 60, and 120 minutes at room temperature before TEG analysis. Coagulation was initiated by adding 20 μL of CaCl₂ to 330 μL of blood and 10 μL of diluted recombinant TF for a final dilution of 1:3600. Reaction (R) and clotting (K) times, angle (α), and maximum amplitude (MA) were compared between time points. A preliminary reference interval (minimum–maximum values) was determined using data from all 20 horses after 30 minutes of sample storage. Results: There was a significant effect of storage time on R, K, and α but not MA. Reference intervals were: R, 3.65–6.4 minutes; K, 1.8–5.45 minutes; α, 33.4–66.2°; MA, 41.2–64.1 mm; lysis at 30 minutes post‐MA (LY30), <2.75%; and lysis at 60 minutes post‐MA (LY60), 1.55–9.5%. Conclusions: TEG can be performed on equine citrated blood samples using recombinant human TF to activate clot formation. TEG parameters were significantly affected by storage time, suggesting an incomplete inhibition of coagulation in citrated blood.     

Thromboelastography in healthy horses and horses with inflammatory gastrointestinal disorders and suspected coagulopathies

Objectives – To evaluate the use of citrated recalcified (nonactivated) thromboelastography (TEG) in healthy horses and horses with colitis and suspected coagulopathies. Design – Prospective, observational study conducted between October 2007 and June 2009. Setting – Veterinary Teaching Hospital. Animals – Forty‐five healthy adult horses and 12 sick adult horses with colitis and prolonged prothrombin time (PT) or activated partial thromboplastin time (aPTT). Interventions – None. Measurements and Main Results – Whole blood was collected on admission. Coagulation profile (PT, aPTT, platelet count, and fibrinogen concentration) and citrated recalcified whole blood TEG analysis (R‐time [R], K‐time [K], angle [α], maximum amplitude [MA], G value [G], lysis at 60 min [LY60]) were evaluated. Mean values (SD) for TEG parameters in healthy horses were: R=10.4 (3.1) minutes; K=3.5 (1.2) minutes; α=46.3 (11.0)°; MA=55.6 (5.1) mm; G=6,429 (1,341) dyn/cm², and LY60=5.1 (2.4)%. Mean coefficients of variation for intra‐assay/interindividual variability in healthy horses were: R=4.7%/30.7%, K=4.8%/35.3%, α=4.4%/23.8%, MA=1.4%/9.3%, G=3.4%/20.8%, and LY60=13.1%/47.7%, respectively. Horses with colitis and prolonged PT and/or aPTT had longer mean values for R (P<0.001) and K (P<0.001), narrower mean α (P<0.001), decreased mean MA (P=0.001), and smaller mean G (P=0.02); changes consistent with hypocoagulability. Conclusions – Citrated recalcified (nonactivated) TEG demonstrated changes consistent with hypocoagulability in horses with colitis that had preidentified coagulation abnormalities. This technique has high interindividual variability and low intra‐assay variability. TEG may be useful for detecting hypocoagulable states in horses with colitis and suspected coagulopathies.     

Synovial fluid D-dimer concentration in foals with septic joint disease

Background: Increased synovial fibrinolytic activity (detected by increases in synovial D‐Dimer concentrations) has been observed in different joint diseases in humans and adult horses, presumably in order to minimize fibrin deposition within the joint and thus avoid its detrimental effects. Objective: To investigate fibrinolytic pathway activation in joint sepsis in foals by measuring synovial D‐Dimer concentrations. Animals: Eighteen septic foals with septic joints, 9 septic foals without septic joints, 9 systemically healthy foals with septic joint, and 3 controls are included. Methods: Prospective observational clinical study of foals admitted for septic arthritis. Synovial D‐Dimer concentration and routine synovial fluid analysis were performed. Diagnosis of joint sepsis was made whenever synovial total nucleated cell count was >30,000 cells/μL, synovial total protein >4 g/dL, and neutrophil percentage of >80%, or synovial fluid culture resulted positive. Results were compared among groups by general lineal models. Results: Synovial D‐Dimer concentration was significantly (P < .001) higher in the foals with septic joints compared with foals without joint disease (P < .001). Conclusions and Clinical Importance: Septic joint disease is associated with a marked increase of synovial D‐Dimer concentration (marked activation of the fibrinolytic activity) within the affected joint. Although further studies are needed, the measurement of synovial D‐Dimer concentration may be considered a complementary diagnostic marker of septic joint disease.     

Coagulation profiles of healthy Andalusian donkeys are different than those of healthy horses

Background: Coagulation disorders are frequently diagnosed, especially in hospitalized equidae, and result in increased morbidity and mortality. However, hemostatic reference intervals have not been established for donkeys yet. Objectives: To determine whether the most common coagulation parameters used in equine practice are different between healthy donkeys and horses. Animals: Thirty‐eight healthy donkeys and 29 healthy horses. Methods: Blood samples were collected to assess both coagulation and fibrinolytic systems by determination of platelet count, fibrinogen concentration, clotting times (prothrombin time [PT] and activated partial thromboplastin time [aPTT]), fibrin degradation products (FDP) and D‐Dimer concentrations. Results: PT and aPTT in donkeys were significantly (P < .05) shorter than those of horses. In contrast, FDP and D‐Dimer concentrations were significantly (P < .05) higher in donkeys than in horses. Conclusions and Clinical Importance: The coagulation parameters most commonly determined in equine practice are different in donkeys compared with horses. Thus, the use of normal reference ranges reported previously for healthy horses in donkeys might lead to a misdiagnosis of coagulopathy in healthy donkeys, and unnecessary treatments in sick donkeys. This is the first report of normal coagulation profile results in donkeys, and further studies are warranted to elucidate the physiological mechanisms of the differences observed between donkeys and horses.     

Examination of hemostatic parameters to detect hypercoagulability in dogs with severe protein‐losing nephropathy

Objective – To identify hemostatic abnormalities in dogs with protein‐losing nephropathies (PLN) that represent risk factors for pathologic thrombosis. Design – Cross‐sectional observational study of client‐owned dogs with PLN, nonprotein losing renal failure (RF), and systemic illness (SI) exclusive of primary renal disease. Setting – Urban University Referral Center. Animals – A total of 29 dogs (n=11 PLN, n=7 RF, n=11 SI) were enrolled between January 2001 and July 2002. Samples were also collected from 20 clinically normal dogs to serve as hemostasis assay controls. Interventions – None. Hemostasis Testing – Citrate anticoagulated blood was collected for point‐of‐care testing with a viscoelastic monitor (thromboelastograph [TEG]) and citrate plasma was prepared for coagulation screening tests and specific assay of the following hemostatic proteins: antiplasmin, antithrombin, D‐dimer, Factor VIII, fibrinogen, plasminogen, protein C, and von Willebrand factor. Results – Dogs with PLN and RF demonstrated TEG abnormalities consistent with hypercoagulability (eg, short clotting time, high clot amplitude) and both groups had significantly lower antithrombin than the SI group. The PLN dogs had significantly higher protein C than either the RF or SI group. Hyperfibrinogenemia was a consistent finding among all 3 disease groups, and the coagulation index a measure of hypercoagulability derived from TEG parameters, directly correlated with fibrinogen values of all study dogs. Conclusions – Hemostatic abnormalities consistent with systemic hypercoagulability are common in dogs with RF and PLN, however, no prothrombotic factors unique to PLN were identified in our study. The thrombotic tendency of PLN may therefore involve parameters we did not directly assess such as platelet reactivity, fibrinolysis, perturbations in blood flow, and/or endothelial dysfunction. High protein C is a novel finding in PLN dogs of unknown clinical relevance.     

Absence of equid herpesvirus-1 reactivation and viremia in hospitalized critically ill horses

Background: Equid herpesvirus‐1 (EHV‐1) reactivation and shedding can occur in latently infected, asymptomatic animals. Risk factors for reactivation include stress and illness. The risk of asymptomatic shedding in hospitalized, critically ill horses with acute abdominal disease is unknown. This information is important to assess the need for additional biosecurity protocols to prevent spread of EHV‐1 in hospitalized critically ill horses with acute abdominal disorders. Objectives: To determine the frequency of reactivation and nasal shedding of EHV‐1 in hospitalized critically ill horses. Animals: One hundred twenty‐four client‐owned horses admitted to the Veterinary Teaching Hospital with acute abdominal disorders were included in the study. Methods: Cross‐sectional study examining the risk of reactivation of EHV‐1 in horses admitted with acute, severe, gastrointestinal disease. Whole blood and nasal secretions were collected throughout hospitalization. In addition, mandibular lymph nodes were collected from 9 study horses and 26 other Michigan horses. All samples were tested for the presence of EHV‐1 nucleic acid by real‐time PCR assays targeting the glycoprotein B gene and the polymerase (ORF 30) gene. Results: One hundred and twenty‐four horses met the inclusion criteria. None of the samples were positive for EHV‐1 DNA. Conclusion and Clinical Importance: These results suggest that nasal shedding and viremia of EHV‐1 in hospitalized critically ill horses with acute abdominal disorder is extremely rare. Implementation of additional biosecurity protocols to limit aerosol spread of EHV‐1 among horses with acute abdominal disease and other hospitalized horses is not necessary.     

The influence of inflammation and hematocrit on clot strength in canine thromboelastographic hypercoagulability

Objective

To investigate parameters causing canine thromboelastographic hypercoagulability and to investigate whether thromboelastography (TEG) with Cytochalasin D (Cyt D) added is related to parameters of platelet activity.

Design

Prospective observational study on hemostatic and inflammatory parameters. Data were collected between November 2012 and July 2013.

Setting

University teaching hospital.

Animals

Twenty‐eight dogs suffering from diseases predisposing to thrombosis and 19 clinically healthy dogs. Diseased dogs were enrolled if they fulfilled inclusion criteria regarding age, size, informed client consent, and obtained a diagnosis of a disease that has been associated with thrombosis or hypercoagulability.

Interventions

None.

Measurements and Main Results

Parameters of coagulation and anticoagulation, fibrinolysis, and antifibrinolysis, platelet activity, inflammation, platelet count, and hematocrit were measured using CBC, TEG, platelet aggregation on multiplate, platelet activity on flow cytometry, and hemostatic and inflammatory markers on plasma and serum analyses. ANOVA and multilinear regression analyses indicated that especially hematocrit and the inflammatory parameters C‐reactive protein and interleukin‐8 showed best association with overall clot strength in diseased dogs with hypercoagulable TEG tracings. Ratios presumed to reflect platelet contribution to the TEG tracing obtained in TEG analyses with Cyt D were related especially with hematocrit and P‐selectin expression of platelets measured after γ‐Thrombin activation on flow cytometry.

Conclusion

Overall clot strength in TEG analyses of the hypercoagulable dogs included in the present study appears to be primarily associated with inflammation as well as hematocrit. Furthermore, the ratio between standard TEG analyses and TEG analyses with Cyt D may reflect some degree of platelet activity.     

Meniscal Mineralization in Domestic Cats

Objective: To (1) determine prevalence of radiographically detectable meniscal mineralization in domestic cats and (2) to evaluate the association between meniscal mineralization and degenerative joint disease (DJD). Study Design: Prospective study. Animals: Client‐owned cats (n=100) and 30 feline cadavers. Methods: Randomly selected client‐owned cats were used to determine the prevalence of meniscal mineralization. Stifles from feline cadavers were used to evaluate the relationship between meniscal mineralization (using high‐resolution X‐ray), radiographic DJD, and cartilage damage. Menisci were evaluated histologically. Results: Forty‐six percent of the client‐owned cats had meniscal mineralization detected in 1 or both stifles. Pain scores were not significantly different between stifles with meniscal mineralization and those with no radiographic pathology (P=.38). Thirty‐four of 57 cadaver stifles had meniscal mineralization, which was always located in the cranial horn of the medial meniscus. Percentage mineralization of the menisci was significantly correlated with the cartilage damage score of the medial femoral (r²=0.6; P<.0001) and tibial (r²=0.5; P<.0001) condyles as well as with the total joint cartilage damage (r²=0.36; P<.0001) score and DJD score (r²=0.8; P<.0001). Conclusion: Meniscal mineralization is a common condition in domestic cats and seems to indicate medial compartment DJD. Clinical Relevance: Clinical significance of meniscal mineralization is uncertain. Further work is needed to determine if the meniscal mineralization is a cause, or a consequence of joint degeneration.     

Influence of subclinical inflammatory airway disease on equine respiratory function evaluated by impulse oscillometry

Reasons for performing study: Inflammatory airway disease (IAD) is a nonseptic condition of the lower respiratory tract. Its negative impact on respiratory function has previously been described using either forced expiration or forced oscillations techniques. However, sedation or drug‐induced bronchoconstriction were usually required. The impulse oscillometry system (IOS) is a noninvasive and sensitive respiratory function test validated in horses, which could be useful to evaluate IAD‐affected horses without further procedures. Objectives: To determine the sensitivity of IOS in detecting alterations of the respiratory function in subclinically IAD‐affected horses without inducing bronchoprovocation and to characterise their respiratory impedance according to frequency for each respiratory phase. Methods: Pulmonary function was evaluated at rest by IOS in 34 Standardbred trotters. According to the cytology of bronchoalveolar lavage fluid (BALF), 19 horses were defined as IAD‐affected and 15 horses were used as control (CTL). Total respiratory resistance (Rrs) and reactance (Xrs) from 1–20 Hz as well as their inspiratory and expiratory components were compared between groups. Results: A significant increase of Rrs at the lower frequencies (R_{1–10 Hz}) as well as a significant decrease of Xrs beyond 5 Hz (X_{5–20 Hz}) was observed in IAD compared to CTL horses. IOS‐data was also significantly different between inspiration and expiration in IAD‐affected horses. In the whole population, both BALF eosinophil and mast cell counts were significantly correlated with IOS measurements. Conclusions: Functional respiratory impairment may be measured, even in the absence of clinical signs of disease. In IAD‐affected horses, the different parameters of respiratory function (Rrs or Xrs) may vary depending on the inflammatory cell profiles represented in BALF. Potential relevance: Impulse oscillometry could be used in a routine clinical setting as a noninvasive method for early detection of subclinical respiratory disease and of the results of treatment in horses.     

Hypercoagulability in dogs with protein-losing enteropathy

Background: Dogs with protein‐losing enteropathy (PLE) have previously been reported to present with thromboembolism; however, the prevalence and pathogenesis of hypercoagulability in dogs with PLE have not been investigated so far. Hypothesis: Dogs with PLE are hypercoagulable compared with healthy control dogs. Animals: Fifteen dogs with PLE. Thirty healthy dogs served as controls (HC). Methods: A prospective study was performed including 15 dogs with PLE. All dogs were scored using the canine chronic enteropathy activity index (CCECAI). Thromboelastography (TEG) and other measures of coagulation were evaluated. Recalcified, unactivated TEG was performed and reaction time (R), kinetic time (K), alpha angle (α), and maximum amplitude (M_A) values were recorded. Nine dogs were reassessed after initiation of immunosuppressive treatment. Results: All dogs with PLE in the study were hypercoagulable with decreased R (PLE: median 7.8, range [2.4–11.2]; HC: 14.1 [9.1–20.3]), decreased K (PLE: 2.5 [0.8–5.2]; HC: 8.25 [4.3–13.1]), increased α (PLE: 56.7 [38.5–78.3]; HC: 25.6 [17–42.4]), and increased M_A (PLE: 68.2 [54.1–76.7]; HC: 44.1, [33.5–49]) (all P < .001). Median antithrombin (AT) concentration was borderline low in PLE dogs; however, mean serum albumin concentration was severely decreased (mean 1.67 g/dL ± 5.1, reference range 2.8–3.5 g/dL). Despite a significant improvement in serum albumin and CCECAI, all 9 dogs with PLE were hypercoagulable at re‐examination. Conclusions and Clinical Importance: The hypercoagulable state in dogs with PLE cannot be solely attributed to loss of AT. Despite good clinical response to treatment, dogs remained hypercoagulable and could therefore be predisposed to thromboembolic complications.     

Effects of zinc-L-carnosine and vitamin E on aspirin-induced gastroduodenal injury in dogs

Background: Nonsteroidal anti‐inflammatory drugs frequently cause gastrointestinal (GI) injury. Zinc‐l ‐carnosine has antioxidant, anti‐inflammatory, mucosal protective, and healing properties in rodent models and in some human studies of GI injury. Hypothesis: The combination of zinc‐l ‐carnosine and vitamin E attenuates aspirin‐induced gastroduodenal mucosal injury. Animals: Eighteen healthy random‐source Foxhound dogs. Methods: In this randomized, double‐blinded, placebo‐controlled study dogs were treated with placebo (n = 6; 0X group), 30 mg/30 IU (n = 6; 1X group), or 60 mg/60 IU (n = 6; 2X group) zinc‐l ‐carnosine/vitamin E orally every 12 hours for 35 days. Between Day 7 and 35, GI mucosal lesions were induced with aspirin (25 mg/kg PO q8h). Mucosal injury lesions (hemorrhage, erosion, and ulcer) were assessed by gastroduodenoscopy on Days 14, 21, and 35 with a 12‐point scoring scale. Results: At baseline (Day ?1) gastroscopy scores were not significantly different between groups (mean ± SD: 0X, 4.4 ± 0.8; group 1X, 4.4 ± 0.6; group 2X, 4.2 ± 0.3; P= .55). Gastroscopy scores increased significantly in all groups between Day ?1 and Days 14, 21, and 35 (P < .0001). On Day 35, gastroscopy scores were 29.2 ± 5.2 (0X), 27.3 ± 3.7 (1X), and 28.6 ± 3.3 (2X). Mean gastroscopy scores were not significantly different among treatment groups on any of the days (P= .61). Conclusions and Clinical Importance: Administration of the combination of zinc‐l ‐carnosine and vitamin E at 1X or 2X dosing did not attenuate aspirin‐induced gastroduodenal mucosal injury.     

Thromboelastography in healthy, sick non-septic and septic neonatal foals

Objectives To evaluate citrated recalcified thromboelastography (TEG) in healthy newborn foals, and to determine intra‐assay, inter‐individual and intra‐individual (at 12 h, 24 h and 7 days after birth) variations. Additionally, to compare TEG variables, haematological values and conventional coagulation profiles from healthy, sick non‐septic, and septic foals. Design Prospective study. Methods The study group comprised 18 healthy, 15 sick non‐septic and 17 septic foals. Two citrated (3.2%; 1 : 9 anticoagulant : blood ratio) blood samples were submitted for haemostatic evaluation using a TEG analyser and conventional coagulation profile. TEG values (R time (R), K time (K), angle (α), maximum amplitude (MA) and G value (G)), complete blood count (CBC) and conventional coagulation profile (prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen concentration (Fib) and antithrombin (AT)) were evaluated. Signalment, presenting complaint, sepsis scores, blood culture results and outcome were taken from the medical records of the sick foals. Results Mean values ± SD for TEG variables in healthy neonatal foals were: R = 11.82 ± 5.35 min, K = 3.06 ± 1.34 min, α= 51.19 ± 12.66 degrees, MA = 55.06 ± 6.67 mm and G = 6361 ± 1700 dyn/cm². Mean coefficients of variation for intra‐assay/inter‐individual/intra‐individual in healthy foals were: R = 3.5/45.2/43.1%; K = 5.3/58.7/28.7%; α= 1.5/24.7/11.9%; MA = 0.3/12.1/6.1%; G = 1.6/26.7/14.7%. Septic foals had significantly greater α, MA and G values than sick non‐septic foals, and significantly greater MA and G than healthy foals, changes that are consistent with hypercoagulability. Weak correlations were detected between TEG variables and haematological or haemostatic values. Conclusions TEG could be used to provide additional information about the haemostatic system in equine neonates.     

Characterization of the inflammatory infiltrate and cytokine expression in the skin of horses with recurrent urticaria

Background – Recurrent urticaria (RU) is a common skin disease of horses, but little is known about its pathogenesis. Hypothesis/Objective – The aim of this study was to characterize the inflammatory cell infiltrate and cytokine expression pattern in the skin of horses with RU. Animals – Biopsies of lesional and nonlesional skin of horses with RU (n = 8) and of skin from healthy control horses (n = 8) were evaluated. Methods – The inflammatory cell infiltrate was analysed by routine histology. Immunohistochemistry was used to identify T cells (CD3), B cells (CD79), macrophages (MAC387) and mast cells (tryptase). Expression of T‐helper 2 cytokines (interleukins IL‐4, IL‐5 and IL‐13), a T‐helper 1 cytokine (interferon‐γ), IL‐4 receptor α and thymic stromal lymphopoietin was assessed by quantitative RT‐PCR. Results – In subepidermal lesional skin of RU‐affected horses, increased numbers of eosinophils (P ≤ 0.01), CD79‐positive (P ≤ 0.01), MAC387‐positive (P ≤ 0.01) and tryptase‐positive cells (P ≤ 0.05) were found compared with healthy horses. Subepidermal lesional skin of RU‐affected horses contained more eosinophils (P ≤ 0.05) and tryptase‐positive cells (P ≤ 0.05) compared with nonlesional skin. There was no significant difference in infiltrating cells between nonlesional skin and skin of healthy horses. Expression of IL‐4 (P ≤ 0.01), IL‐13 (P ≤ 0.05), thymic stromal lymphopoietin (P ≤ 0.05) and IL‐4 receptor α (P ≤ 0.05) was increased in lesional skin of RU‐affected horses compared with control horses. Expression of IL‐4 was higher (P ≤ 0.05) in lesional compared with nonlesional RU skin. Conclusions and clinical importance – Analysis of cytokine expression and inflammatory infiltrate suggests that T‐helper 2 cytokines, eosinophils, mast cells and presumptive macrophages play a role in the pathogenesis of equine RU.     

Cartilage-derived retinoic acid-sensitive protein in equine synovial fluid from healthy and diseased joints

Reasons for performing study: More sensitive and specific diagnostic methods for early detection of changes in the joint cartilage are needed. Cartilage‐derived retinoic acid‐sensitive protein (CD‐RAP) is a potential marker of cartilage synthesis and regeneration. This is the first study on equine CD‐RAP. Objectives: To evaluate the ability of a commercially available human sandwich ELISA assay to detect equine CD‐RAP in synovial fluid from healthy and diseased joints. Methods: Synovial fluid was collected from 28 horses with no signs of joint disease and from 5 with induced inflammatory arthritis. CD‐RAP concentrations were measured using a human CD‐RAP ELISA. Intra‐ and interassay imprecision of the assay were evaluated by multiple measurements on pools of equine synovial fluid. Assay inaccuracy was determined by linearity under dilution. Results: The assay showed moderate to large intra‐ and interassay variation when applied to equine synovial fluid. Equine CD‐RAP was detected in synovial fluid from healthy horses ranged at 8.2–52 ng/ml. Repeated arthrocentesis (after injection of isotonic saline), age, joint or gender did not significantly affect CD‐RAP concentrations. Twelve hours after intra‐articular injection of lipopolysaccharide, concentrations of CD‐RAP were significantly lower than after injection of isotonic saline and remained significantly lower until the end of the study at 144 h. Conclusion and potential relevance: The assay is suitable for longitudinal monitoring of CD‐RAP concentration in individual horses. Disease significantly influenced CD‐RAP levels. Similar to previous results obtained in man, CD‐RAP seems to be a marker of cartilage synthesis and/or regeneration in horses.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.